14 results about "Cell dissociation" patented technology

This invention provides a method for propagating pluripotent stem cells (PSCs) by suspension culture in a bioreactor. [Solution] The method comprises the following steps: (i) adding a ROCK inhibitor (ROCKi) to pluripotent stem cells cultured in suspension in a bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding a sufficient excess volume of culture medium to reduce the concentration of the cell dissociation agent to a concentration at which cell aggregates can be re-formed; and (iv) culturing the mixture obtained in step (iii) under appropriate conditions that allow for the proliferation of PSCs.

This invention discloses a multi-enzyme preparation for industrial cell dissociation, its usage method, and applications, belonging to the field of seed cell extraction technology for industrial meat culture. The invention employs a multi-enzyme preparation composed of component 1 and component 2; component 1 includes neutral protease, hyaluronidase, and elastase; component 2 includes matrix metalloproteinase, papain, and plasmin. The invention also discloses the preparation method and applications of the multi-enzyme preparation. The rational enzyme ratio and optimized treatment conditions of this invention help protect cell membrane integrity, maintain high cell activity, and significantly improve dissociation efficiency and cell recovery. The compound enzyme preparation is highly adaptable, applicable to various tissue types and dissociation requirements, and facilitates standardized and large-scale production. Combined with automated dissociation equipment, the compound enzyme preparation enables efficient and stable industrial operation, providing important technical support and broad application prospects for the cultured meat industry, tissue engineering, and cell therapy.

A method for culturing primary gynecological tumor cells and culture medium used therein. The method includes using mild cell dissociation reagents to treat a gynecological solid tumor tissue, to ensure the vitality of tumor cells in the tissue to the greatest extent; preparing a special serum-free medium, and using a suspension culture system to culture solid tumor cells derived from a gynecological tumor in vitro to ensure the normal expansion of tumor cells and eliminate interference from normal cells to the greatest extent. The primary gynecological tumor cell culture obtained by the method of the present invention can be used for various cell-based in vitro experiments, second-generation sequencing, construction of animal models, construction of cell lines and the like. It is foreseeable that this culture method has broad application prospects in the fields of gynecological tumor research and clinical diagnosis and treatment.

The invention discloses an efficient universal mouse pancreatic tissue single-cell suspension preparation method, and belongs to the technical field of cell biology. Pancreas are rich in digestive enzymes, so that the treatment difficulty of in-vitro cells is high, and a high-quality single-cell suspension is difficult to prepare by a traditional method. The method comprises the steps of reagent preparation, tissue sampling, tissue dissociation, red cracking and cleaning, AOPI dyeing quality inspection and the like. Components and concentrations of a tissue enzymatic hydrolysate, a cell cleaning solution and a cell suspension are determined in reagent preparation; the acquisition method of the pancreatic tissue of the mouse is defined by tissue sampling; the tissue dissociation adopts a two-round digestion mode, and the cut tissue is subjected to enzymolysis by using a compound enzyme solution under specific conditions; the cell precipitates are treated through red cracking and cleaning; aOPI dyeing quality inspection is used for analyzing cell count and motility rate. According to the method, the culture medium containing specific components and the compound enzyme liquid are creatively used, the problem that pancreas single cells are difficult to dissociate can be effectively solved, the single-cell suspension with high motility rate, high purity and low adhesion is obtained, and the requirements of downstream single-cell experiments are met.

An automated system for anticancer drug sensitivity testing, according to an embodiment of the present invention, may comprise: a chopper configured to mince patient tissue; a cell dissociation device configured to apply a cell dissociation enzyme to the patient tissue minced by the chopper; a centrifuge configured to centrifuge cells to which the cell dissociation enzyme has been applied by the cell dissociation device; a liquid handler configured to mix the centrifuged cells with extracellular matrix to thereby generate a mixed sample and to inject the mixed sample into a nozzle tip; a spotter configured to dispense the mixed sample onto a pillar by using the nozzle tip; an automatic incubator configured to culture the mixed sample to generate cell organoids; a scanner configured to generate an image by scanning the cell organoids at high speed; and a multi-joint robot configured to move at least one of the patient tissue, the cells, and the mixed sample between at least some of the chopper, the cell dissociation device, the centrifuge, the liquid handler, the spotter, the automatic incubator, and the scanner. Thus, the entire process of cell extraction from tissue isolated from a patient to organoid culture, drug testing, and scanning can be performed fully automatically.

The application discloses a kind of high efficiency gentle mammal blastocyst single cell dissociation method.The method includes the following steps: (1) zona pellucida removal: the blastocyst is placed in calcium magnesium table type liquid, and zona pellucida is completely removed by dissolving and physical blowing operation;(2) enzyme digestion incubation: the blastocyst after step (1) is handled and is transferred to 35-37 ℃ pre-equilibrated complex dissociation solution and incubated for 15-30 minutes, during which intermittent blowing is carried out, until the blastocyst is completely dissociated into single cell;(3) washing: the single cell obtained by dissociation is washed using dPBS, and the residual dissociation solution is removed.The method of the application has high efficiency, high activity and simplified operation.The dissociation process is simplified from 3-4 steps to two steps (zona pellucida dissolution+enzyme digestion incubation), the total time is shortened from more than 90 minutes to less than 30 minutes, the dissociation efficiency is significantly improved;Cell viability is increased from about 70% to more than 90%, the cell junction is gently lysed, and cell damage is reduced.

The invention relates to the technical field of non-enzymatic cell dissociation, in particular to a non-enzymatic adipose tissue dissociation method and device based on hydrodynamic shearing, and the method comprises the following steps: step 1, obtaining dissociation indexes, and obtaining an optimal process parameter combination I meeting the dissociation indexes through a model based on the dissociation indexes; step 2, establishing a reference process based on the optimal process parameter combination I; step 3, judging whether a decision is input or not, if so, entering step 4, and otherwise, performing a dissociation process based on a reference process; and step 4, optimizing the reference process based on decision input to obtain an optimal process parameter combination 2, and performing a dissociation process based on the optimal process parameter combination 2.

The invention discloses a mouse epididymal body extraction method which comprises the following steps: S1, digesting mouse epididymal tissues: putting separated mouse epididymis into currently prepared mouse sperm insemination liquid, cutting into pieces, discarding sperms, and adding cell dissociation liquid to form single-cell suspension; s2, purification of mouse epididymis exosomes: filtering the single-cell suspension, and purifying, eluting and extracting the exosomes through an exoEasy Maxi Kit kit; s3, concentration of the epididymis bodies: performing ultracentrifugation on the eluent of the exosomes to concentrate the exosomes, discarding the supernatant to obtain precipitates, namely the high-purity epididymis bodies, and performing identification after subsequent resuspension. The method has the advantage that the mouse epididymis body with high purity, high yield and complete structure can be obtained.

The present application relates to the field of biotechnology, in particular to human bladder tissue digestion solution and application thereof.The digestion method of the present application can dissociate human bladder tissue cells while keeping the cell viability at more than 90%, and the amount of harvested cells is also obviously improved.Sufficient cell amount is the prerequisite for optimization treatment such as red blood cell lysis and dead cell removal.The single cell suspension obtained by the method has high viability, low clumping rate, high nucleated rate and less impurities, which can provide a basis for obtaining high-quality gene expression data in human bladder tissue single cell sequencing experiment.The present application can quickly obtain human bladder tissue primary cell suspension (sufficient cell amount, high viability, low clumping rate and less impurities) meeting the requirements of single cell experiment within 2-4 hours.

The present invention relates to new plasma or new platelet-rich plasma preparations, new cell dissociation methods, new cell associations or compositions, a method of preparation thereof, a use thereof, devices for the preparation thereof and preparations containing such a platelet-rich plasma preparation and cell associations or compositions. Specifically, the invention provides plasma or platelet-rich plasma alone or in cell composition preparations for use in tissue regeneration and bone regeneration and pain reduction.

This invention provides a kit, method, and single-cell library construction method for the dissociation of marine animal tissues. The dissociation method includes: S1) dissociating marine animal tissue samples at 8℃~12℃ using a cell dissociation buffer to obtain a single-cell suspension; S2) centrifuging the single-cell suspension and washing the single-cell precipitate obtained after centrifugation with a cell resuspension buffer to obtain single cells from the marine animal tissue sample; wherein, the cell dissociation buffer comprises a protease; the protease is Bacillus subtilis protease A; the concentration of sodium ions in the cell resuspension buffer is 300-800mM, and the concentration of potassium ions is 5-20mM.

A method for collecting cells from a bioreactor is provided wherein the bioreactor comprises cells to be collected, at least one inlet and / or outlet, and a culture chamber in which the cells are attached to a matrix. The method comprises providing a wash solution having a cell dissociation reagent (CDR) into the culture chamber; and after providing the washing solution, removing and capturing the detached cells from the culture chamber via the at least one inlet and / or outlet. The cell dissociation agent includes an endonuclease, in particular a deoxyribonuclease, such as deoxyribonuclease I (DNase I). The wash solution may further include a divalent metal ion, such as Mg + 2, Ca + 2, or Mn + 2.

The invention relates to the technical field of cell culture, in particular to a human pluripotent stem cell culture passage dissociation reagent and application thereof. On the basis that sodium ethylene diamine tetracetate is commonly used as a key component of a cell dissociation reagent, a formula is optimized, and the following components are added: amino acid, a surfactant PEG (polyethylene glycol) and glucose, specifically arginine hydrochloride, lysine hydrochloride, sodium aspartate, PEG8000 and glucose. The added components are high in solubility and stable in property, the optimized dissociation reagent is easy and convenient to prepare, the implementation feasibility of the technical scheme is high, low damage to cells is guaranteed in the dissociation process, the cell viability is better maintained, and the quality of cells after passage is improved.

The invention provides cell culture conditions for culturing stem cells, including feeder-free conditions for generating and culturing human induced pluripotent stem cells (iPSCs). More particularly, the invention provides a culture platform that allows long-term culture of pluripotent cells in a feeder-free environment; reprogramming of cells in a feeder-free environment; single-cell dissociation of pluripotent cells; cell sorting of pluripotent cells; maintenance of an undifferentiated status; improved efficiency of reprogramming; and generation of a naïve pluripotent cell.